Embryo-mediated genome editing for accelerated genetic improvement of livestock

doi:10.15302/J-FASE-2019305

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Abstract：

Selecting beneficial DNA variants is the main goal of animal breeding. However, this process is inherently inefficient because each animal only carries a fraction of all desirable variants. Genome editing technology with its ability to directly introduce beneficial sequence variants offers new opportunities to modernize animal breeding by overcoming this biological limitation and accelerating genetic gains. To realize rapid genetic gain, precise edits need to be introduced into genomically-selected embryos, which minimizes the genetic lag. However, embryo-mediated precision editing by homology-directed repair (HDR) mechanisms is currently an inefficient process that often produces mosaic embryos and greatly limits the numbers of available edited embryos. This review provides a summary of genome editing in bovine embryos and proposes an embryo-mediated accelerated breeding scheme that overcomes the present efficiency limitations of HDR editing in bovine embryos. It integrates embryo-based genomic selection with precise multi-editing and uses embryonic cloning with elite edited blastomeres or embryonic pluripotent stem cells to resolve mosaicism, enable multiplex editing and multiply rare elite genotypes. Such a breeding strategy would enable a more targeted, accelerated approach for livestock improvement that allows stacking of beneficial variants, even including novel traits from outside the breeding population, in the most recent elite genetic background, essentially within a single generation.

Keywords animal breeding cattle cloning CRISPR/Cas9 cytoplasmic injection embryo genome editing germline chimaeras HDR livestock improvement TALENs

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	Zachariah MCLEAN
	Björn OBACK
	Götz LAIBLE

引用本文:

Zachariah MCLEAN,Björn OBACK,Götz LAIBLE. Embryo-mediated genome editing for accelerated genetic improvement of livestock[J]. Front. Agr. Sci. Eng. , 2020, 7(2): 148-160.

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https://journal.hep.com.cn/fase/EN/10.15302/J-FASE-2019305 OR https://journal.hep.com.cn/fase/EN/Y2020/V7/I2/148

Fig.1 Soma- and embryo-mediated genome editing approaches in cattle. (a) Somatic cells isolated from an elite sire are genome edited (locus black to gold) and cloned to introduce the modification on the same genetic background. A new elite sire with a higher breeding worth—and carrying the genomic modification—is produced after crossing the cloned sire with an elite dam. (b) For embryo-mediated genome editing, the elite sire and dam are crossed, and the genome editing carried out in the zygote, introducing the genomic modification in the higher breeding worth genetic background in a single generation.

Tab.1 Publications of embryo-mediated genome editing in bovine zygotes

Fig.2 Cytoplasmic injection in a bovine zygote. Image courtesy of Dr. Jingwei Wei (AgResearch, Ruakura Research Centre, New Zealand).

Fig.3 Resolving mosaicism in edited embryos by embryonic cell transfer (ECT) blastomere cloning. After genome editing in the zygote, a mosaic morula can be dissociated and the blastomeres used for ECT. The resulting embryos derived from a single blastomere would then be non-mosaic, which can be confirmed with an embryo biopsy before transfer or cryopreservation.

Fig.4 Multiplex genome editing in embryonic pluripotent stem cells (ePSCs) isolated from a genomically selected elite blastocyst. With high clonogenicity and stable pluripotency, ePSCs would allow sequential multiplex genome editing of multiple loci (loci black to gold), followed by clonal expansion of edited strains.

Fig.5 Embryo-mediated accelerated breeding in cattle. (a) Using proven technologies (solid arrows), precise genomic edits can be introduced in zygotes from elite sires and the best embryos identified by genomic selection (GS). The desired embryonic genotype can then be multiplied by embryonic cell transfer (ECT) cloning and elite offspring generated. (b) A complementary pathway (dashed arrows) is centered around embryonic pluripotent stem cells (ePSCs). ePSCs would be isolated from an elite embryo and numerous genomic modifications introduced. Alternatively, ePSCs would be isolated from embryos multiplied by ECT for subsequent genome editing. Offspring could then be produced by cloning or as a chimaera with absolute transmission of the elite germline.

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